Hepatitis B virus capsid assembly

Abstract

Cell-free translation and assembly systems allow for HBV capsid assembly under cell-free conditions that also mimic the physiological salt and protein concentrations. These hepatitis virus capsid assembly systems utilize the C-terminal domain (CTD) and N-terminal domain (NTD) of the hepatitis capsid protein in capsid assembly. Uses of the system include the identification of potential new therapeutic strategies and screening of potential therapeutic agents which target these domains and modulate capsid assembly and/or disassembly and other functions of these domains in HBV replication and pathogenesis.

What is claimed is: 
     
       1. A method for assembling Hepatitis virus capsids in vitro comprising: transcribing, translating or expressing one or more hepatitis virus capsid or core protein nucleic acids, polynucleotides, oligonucleotides, proteins, peptides, mutants or fragments thereof, which encodes for or comprises a hepatitis virus capsid or core protein C-terminal Domain (CTD) and N-terminal domain (NTD) sufficient to form capsids, in a rabbit reticulocyte lysate (RRL) cell-free system, said RRL cell-free system comprising at least one host factor for controlling a phosphorylation state of a capsid protein, said at least one host factor being Cyclin-dependent kinase 2 (CDK2) or Protein phosphatase 2 (PP2A). 
     
     
       2. The method of  claim 1 , wherein an assembled Hepatitis virus capsid comprises one or more agents encapsulated therein. 
     
     
       3. The method of  claim 2 , wherein the one or more agents comprise: chemotherapeutic agents, anti-virus agents, anti-inflammatory agents, nucleic acids, vectors, expression vectors, polynucleotides, oligonucleotides, proteins, peptides, lipids, lipoproteins, organic molecules, inorganic molecules, synthetic compounds, natural compounds, saccharides, or combinations thereof. 
     
     
       4. The method of  claim 1 , wherein the assembled hepatitis virus capsid is a non-replicating and/or non-infecting hepatitis B virus capsid.